Yoonk Sik Sung Gotina, Lizaveta Kim, Kyu Hyeon Lee, Jung Yeol Seulgi Shin Aziz, Hira Kang, Dong Min Liu, Xiao Hong, Na-Kyeong Lee, Hong-Guen Lee, Jun-Seok Ku, Hyeyeong Jeong, Cherl hyun Pae, Ae Nim Lim, Sungsu Chang, Young-Tae Kim, Yun Kyung 2024-01-12T02:30:07Z 2024-01-12T02:30:07Z 2023-12-14 2024-01 1433-7851 https://pubs.kist.re.kr/handle/201004/75732 The development of a small-molecule probe designed to selectively target neurons would enhance the exploration of intricate neuronal structures and functions. Among such probes, NeuO stands out as the pioneer and has gained significant traction in the field of research. Nevertheless, neither the mechanism behind neuron-selectivity nor the cellular localization has been determined. Here, we introduce NeuM, a derivative of NeuO, designed to target neuronal cell membranes. Furthermore, we elucidate the mechanism behind the selective neuronal membrane trafficking that distinguishes neurons. In an aqueous buffer, NeuM autonomously assembles into micellar structures, leading to the quenching of its fluorescence (Φ=0.001). Upon exposure to neurons, NeuM micelles were selectively internalized into neuronal endosomes via clathrin-mediated endocytosis. Through the endocytic recycling pathway, NeuM micelles integrate into neuronal membrane, dispersing fluorescent NeuM molecules in the membrane (Φ=0.61). Molecular dynamics simulations demonstrated that NeuM, in comparison to NeuO, possesses optimal lipophilicity and molecular length, facilitating its stable incorporation into phospholipid layers. The stable integration of NeuM within neuronal membrane allows the prolonged monitoring of neurons, as well as the visualization of intricate neuronal structures. English John Wiley & Sons Ltd. NeuM: A Neuron-Selective Probe Incorporates into Live Neuronal Membranes via Enhanced Clathrin-Mediated Endocytosis in Primary Neurons Article 10.1002/anie.202312942 1 Angewandte Chemie International Edition, v.63, no.3 Angewandte Chemie International Edition 63 3 N scie scopus 001129182500001 Chemistry, Multidisciplinary Chemistry Article CHARMM GUI MECHANISMS DYE Clathrin-Mediated Endocytosis Endocytic Trafficking Fluorescent Probe Membrane Neuron